Wnt3a promotes osteogenic differentiation of periodontal ligament stem cell and regeneration of alveolar bone in experimental periodontitis
10.3760/cma.j.cn112144-20200611-00334
- VernacularTitle:Wnt3a促进牙周膜干细胞成骨分化及实验性牙周炎牙槽骨再生的研究
- Author:
Song SHEN
1
;
Shengjun SUN
;
Shaohua GE
Author Information
1. 山东大学齐鲁医学院口腔医学院·口腔医院牙周科 山东省口腔组织再生重点实验室 山东省口腔生物材料与组织再生工程实验室，济南 250012
- Keywords:
Wnt proteins;
Periodontitis;
Periodontal ligament stem cells;
Alveolar bone regeneration
- From:
Chinese Journal of Stomatology
2021;56(3):268-275
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effects of Wnt3a on the proliferation, migration and osteogenic differentiation of periodontal ligament stem cell (PDLSC) and to identify the role of Wnt3a in alveolar bone regeneration in mouse experimental periodontitis.Methods:The experiments were conducted by stimulating PDLSC using Wnt3a of 5 different concentrations (0, 20, 100, 200, 500 μg/L) respectively. Cell proliferation was detected by cell-counting assay, cell migration was evaluated by Transwell assay and the expressions of osteogenic related genes collagen Ⅰ (Col-Ⅰ), runt-related transcription factor 2 (Runx2) were examined by real-time quantitative PCR (RT-qPCR). Poly lactic-co-glycolic acid (PLGA)-Wnt3a-hyaluronic acid (HA) hydrogel was injected locally into the gingival sulcus of mice with experimental periodontitis. After 1, 2, 4, and 8 weeks of hydrogel injection, samples of maxillary alveolar bone were obtained. Micro-CT, HE staining and immunohistochemical staining of osteogenesis related markers, such as alkaline phosphatase (ALP), Runx2, osteocalcin (OCN), were used to evaluate alveolar bone regeneration.Results:After 10 d of culture, Wnt3a with concentrations of 20-500 μg/L significantly promoted the proliferation ( P<0.01) and the migration ( P<0.01) of PDLSC. After 21 d of culture, the expression levels of Col-Ⅰ mRNA were 0.96±0.27, 1.90±0.47, 2.18±0.24, 2.32±0.15 and 1.99±0.43 in 5 concentration groups respectively, and the expression levels of Runx2 mRNA were 1.08±0.15, 3.19±0.17, 6.19±0.28, 9.19±0.41 and 5.55±0.06, respectively. Both expressions had significant statistical differences compared with the negative control group ( P<0.05). At 1, 2, 4, and 8 weeks, the Wnt3a hydrogel group had less distance [(497.3±18.2), (455.7±12.5), (401.0±8.5), (362.3±15.5) μm] from the cemento-enamel junction to alveolar bone crest compared with the periodontitis group [(710.3±10.2), (614.0±16.4), (564.3±12.5), (502.3±6.8) μm] ( P<0.01) and weaker periodontal inflammation. Immunohistochemical results showed that the expression levels of bone-related proteins of ALP (0.72±0.01), Runx2 (0.77±0.03) and OCN (0.72±0.07) in the Wnt3a hydrogel group were increased compared with the periodontitis group ( P<0.01). Conclusions:Wnt3a might promote the proliferation, migration and osteogenic differentiation of PDLSC and the alveolar bone regeneration.
